酿酒酵母乙酸耐受性相关的微卫星分子标记筛选

Screening of microsatellite markers associated with acetic acid tolerance of Saccharomyces cerevisiae

摘要:

【目的】筛选与酵母乙酸耐受性状紧密相关的微卫星分子标记。【方法】以两株表型差异菌株YHA和YLA作为亲本构建F2代菌株共计160株，选取15个微卫星位点通过PCR方法在40株子代中扩增产物，利用SPSS 11.5软件分析耐酸性状与微卫星序列间的相关性。【结果】找到3个与乙酸耐受性性状相关的微卫星位点，其中位点14P2与酵母乙酸耐受性状有极显著的正相关性(P<0.01)，15P2和15P3与酵母乙酸耐受性具有显著的负相关性(P<0.01和P<0.05)；此外对于微卫星位点14P2，耐酸亲本YHA在该位点的基因片段(344 bp)在子代耐酸菌株中出现频率达到70.6%，而不耐酸亲本YLA的基因片段(331 bp)在子代不耐酸菌株中出现的频率达91.3%。【结论】微卫星14P2的等位基因在子代菌株中的遗传具有明显的偏好性，该微卫星位点与某种耐酸基因存在一定的连锁遗传，为酵母分子标记辅助育种提供了有价值的遗传标记。

Abstract:

[Objective] To screen microsatellite markers associated with acetic acid tolerance of Saccharomyces cerevisiae. [Methods] One hundred and sixty segregants were obtained by crossing two yeasts (high acetate tolerant YHA and low acetate tolerant YLA) with different phenotypes. The alleles of 15 microsatellite markers were amplified from 40 offspring yeasts and SPSS 11.5 software was used to analysis the relationship between acetic acid tolerance and microsatellite markers. [Results] Three microsatellite loci were identified. Among them marker 14P2 had significant positive correlation with acid tolerance (P<0.01), marker 15P2 and marker 15P3 had significant negative correlation with acid tolerance (P<0.01 and P<0.05). Furthermore, 70.6% of high tolerant individuals contained the marker 14P2 (344 bp) derived from YHA, the high tolerant strain; 91.3% of low tolerant individuals contained the marker 14P2 (331 bp) derived from YLA, the low tolerant strain. [Conclusion] This study indicated that the microsatellite marker 14P2 has the obvious tendency of heredity correlative with genes for acetic acid tolerance, it can be used in molecular marker assisted selection.